Documentation of Indigenous Antiparasitic Practices and Scientific Evaluation of Some Ethnobotanicals for Their Anthelmintic Activity
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DOCUMENTATION OF INDIGENOUS ANTIPARASITIC PRACTICES AND SCIENTIFIC EVALUATION OF SOME ETHNOBOTANICALS FOR THEIR ANTHELMINTIC ACTIVITY By Nadeem Badar Reg. # 91-ag-750 A Thesis Submitted in Partial Fulfillment of the Requirement for the Degree of DOCTOR OF PHILOSOPHY IN PARASITOLOGY FACULTY OF VETERINARY SCIENCE UNIVERSITY OF AGRICULTURE FAISALABAD, PAKISTAN 2011 DECLARATION I hereby declare that contents of the thesis, “Documentation of indigenous antiparasitic practices and scientific evaluation of some ethnobotanicals for their anthelmintic activity” are product of my own research and no part has been copied from any published source (except the references, some standard mathematical or genetic models/ equations/protocols etc.). I further declare that this work has not been submitted for award of any other diploma/degree. The university may take action if the information provided is found inaccurate at any stage. (In case of any default the scholar will be proceeded against as per HEC plagiarism policy). _____________________ Signature of the Student Name: Nadeem Badar Reg. No. 91-ag-750 ACKNOWLEDGEMENT First of all I am thankful to my Allah Who enabled me to collect and to contribute my findings in the existing scientific knowledge. Then I record my deep sense of gratitude and appreciation to Prof. Dr. Zafar Iqbal, Department of Parasitology, University of Agriculture, Faisalabad for his constant encouragement and inspiring guidance throughout my postgraduate period. I have no appropriate words that fully convey the sense of immense thanks and deep gratitude that I owe to my respectable committee members Prof. Dr. Muhammad Nisar Khan, Chairman, Department of Parasitology, University of Agriculture, Faisalabad and Prof. Dr. Muhammad Shoaib Akhtar for their kind supervision and affections. I am grateful to the Lab. Attendants and those who co-operated with me during data collection. I can not forget the prayers and encouragement of my affectionate parents who always supported me throughout my life and I am also thankful to the higher education commission (HEC) of Pakistan for partial funding of this project. Nadeem Badar CONTENTS Chapter No. Contents Page No. LIST OF TABLES i FIGURE iii APPENDIX iii 1 INTRODUCTION 1 2 REVIEW OF LITERATURE 3 3 MATERIALS AND METHODS 18 4 RESULTS 27 5 DISCUSSION 54 6 SUMMARY 73 7 REFERENCES 75 LIST OF TABLES Table No. Content Page No. 1 Plants used as anthelmintics 4 2 Plants used as antiprotozoals 9 3 Plants used as insecticides 12 4 Plants used as acaricides 14 5 Tests/assays for scientific activity evaluation of different 16 ethnobotanicals 6 Plants selected for anthelmintic activity 22 7 Layout plan for crude powder and crude aqueous methanolic extract of 25 Acacia nilotica (Bark & Leaves), Vitex negundo (seeds) and Arundo donax (leaves) given to different groups of sheep naturally infected with mixed species of gastrointestinal nematodes 8 Layout plan for crude powder and crude aqueous methanolic extract of 25 Amomum subulatum (Fruit) given to different groups of sheep naturally infected with mixed species of gastrointestinal nematodes 9 Layout plan for crude powder and crude aqueous methanolic extract of 26 Areca catechu (seeds) and Ferula assa-foetida (latex) given to different groups of sheep naturally infected with mixed species of gastrointestinal nematodes 10 Botanical, local and English names of the plants documented from 27 district Jhang (Punjab, Pakistan) for their use in ethnoveterinary medicine 11 Name of plants, representing families and frequency of their usage in 28 ethnoveterinary medicine in district Jhang (Punjab, Pakistan) 12 Top ten most frequently reported plants for their usage in 29 ethnoveterinary medicine in Jhang (Punjab, Pakistan) 13 Inventory of plants used for the treatment of different diseases/ 30 conditions of livestock reported by the local respondents in Jhang (Punjab, Pakistan) 14 Number and nature of EVM practices for different diseases/conditions 35 documented from Jhang (Punjab, Pakistan) 15 Plants, diversity of their usage and contribution in total number of 36 EVM practices for different diseases/conditions of livestock in Jhang (Punjab, Pakistan) 16 In vitro effect of crude aqueous methanol extracts of different plants on 39 survival of H. contortus of sheep in comparison with Levamisole 17 In vitro effect of different fractions of crude aqueous methanol extracts 40 of Ferula assa-foetida on survival of H. contortus of sheep in comparison with Levamisole i LIST OF TABLES…continued Table No. Content Page No. 18 In vitro effect of different fractions of crude aqueous methanol extracts 41 of Acacia nilotica bark on survival of H. contortus of sheep in comparison with Levamisole 19 In vitro effect of different fractions of crude aqueous methanol extracts 42 of Areca catechu on survival of H. contortus of sheep in comparison with Levamisole 20 In vitro effect of different fractions of crude aqueous methanol extracts 43 of Amomum subulatum on survival of H. contortus of sheep in comparison with Levamisole 21 In vitro effect of different fractions of crude aqueous methanol extracts 44 of Vitex negundo on survival of H. contortus of sheep in comparison with Levamisole 22 In vitro effect of different fractions of crude aqueous methanol extracts 45 of Acacia nilotica leaves on survival of H. contortus of sheep in comparison with Levamisole 23 In vitro effect of different fractions of crude aqueous methanol extracts 46 of Arundo donax on survival of H. contortus of sheep in comparison with Levamisole 24 Ranking of efficacy of different fractions of plants against H. contortus 46 25 Percent egg hatch and LC50 of crude aqueous methanol extracts and 47 fractions of different plants 26 Top 10 highly ovicidal plant crude aqueous methanol extracts and/or 48 fractions in comparison with control 27 Regression values and correlation of regression of the effect of 49 different plants on egg hatching 28 Top 10 plant crude aqueous methanol extracts and/or fractions having 50 best dose dependant ovicidal effects 29 Effect of crude powder and crude methanol extracts of different plants 51 on eggs per gram of feces (Mean±SEM) in sheep naturally infected with mixed species of gastrointestinal nematodes in comparison with untreated and levamisole treated animals 30 Efficacy ranking based on reduction in faucal egg counts as on day 12 52 post-treatment (PT) of crude powder and crude methanol extract of different plants 31 Ranking of efficacy of different CAME/fractions of plants based on 53 three tests employed in the study ii FIGURE Fig. No. Content Page No. 1 Map of Pakistan showing the study area 19 APPENDIX Appendix Content Page No. No. 1 List of private Livestock farms included in the study 25 iii Chapter # 1 Introduction Livestock production depends on the feed supplies, good health coverage and appropriate animal husbandry practices. Many climatic and casual factors are experiencing high degree of parasitic attack and plague on them resulting in their decreased longevity, survivorship, infertility and productivity. The productive and reproductive potential of domestic livestock is adversely affected because of clinical and sub-clinical infections. All the ailments afflicting animals are counterproductive. Gastrointestinal helminthiasis (especially nematodosis) is, however, of high economic significance in view of its insidious nature and easy transmissibility due to under feeding, availability of a wide variety of hosts, vectors, inadequate/low level of awareness and animal health cover. For centuries, most of the human and animal population in Indo-Pakistan subcontinent has relied on a system of traditional medicine. The traditional medicine system prevailing in this area is also termed as Eastern Medicine, Unani Medicine, Islamic Medicine or Ethno-medicine. Ethno-veterinary medicine (EVM) is the traditional system of maintaining animal health and curing diseases of animals that is based on folk beliefs and traditional knowledge, skills, methods and practices (Mathias-Mundy and McCorkle, 1989). EVM is a traditional system that local people, through trial-and-error and also deliberate experimentation, developed to keep their animals healthy and productive. Though, this old method of treatment has been replaced by allopathy in some parts of globe (Tabuti et al., 2003), yet it is contributing significantly in improving the animal health in developing countries. It can play a very important role in sustainable veterinary medicine in modern world (Lin et al., 2003), especially in Indian subcontinent due to various benefits associated with this method of treatment, e.g. lower cost, greater accessibility and apparent efficacy (Mwale et al., 2005). Tradition of EVM is very old in Indo-Pakistan region and it is evident from its documentation in the form of Rig-veda (from 4500 to 1600 BC) and Ayurveda from 2500 to 600 BC (from Zaman and Khan, 1972; Somvanshi, 2006). Old books and scriptures of 2350 BC like; Agni Purana, Skanda Purana, Matsya Purana, Devi Purana, Garuda Purana and Linga Purana describes the documentation of phytotherapy for treatment of different diseases of animals (Somvanshi, 2006). Majority of the EVM surveys and validation studies indicate much wider and effective use of plants as anthelmintics compared with other diseases/conditions (Iqbal et al., 2004, 2005, 2006, 2006a,b,c; Jabbar et al., 2007; Hussain et al., 2008). The tremendous use of plants as anthelmintics for the treatment of helminthiasis is attributed to its high prevalence and heavy 1 production